VEGF基因修饰兔骨髓间充质干细胞的研究**☆

背景：血管内皮生长因子的应用是组织工程组织血管化的有效手段。但是，血管内皮生长因子价格昂贵，并且半衰期非常短，很难在体内保持有效的作用浓度。故本实验将其转入组织工程的种子细胞——骨髓间充质干细胞中，使骨髓间充质干细胞能够有效地分泌血管内皮生长因子，从而达到促进血管生成的目的。 目的：探讨应用人血管内皮生长因子165进行基因修饰的可行性，为血管化组织工程组织的构建及缺血性疾病的治疗奠定实验基础。 设计：观察对比实验。 单位：吉林大学第一医院和吉林大学再生医学科学研究所。 材料：实验于2003-06/2004-08 在吉林大学再生医学科学研究所吉林大学重点实验室（生物安全实验室二级）完成。健康新西兰大耳白兔由吉林大学实验动物中心提供。4.0~5.0 月龄，体质量215~315 kg，雌雄兼用，实验过程中对动物处置均在麻醉状态、无菌条件下完成，符合动物伦理学标准。实验药品及试剂：Ham F12 培养基( GIBCO公司)，噻唑蓝(MTT) ( Sigma 公司)，pLXSN-KDRp-VEGF165 与pcDNA3.0 载体由本实验室自备, ELISA 检测试剂盒(深圳晶美生物公司)，感受态大肠杆菌DH5α、限制性内切酶BamH I、Xho I、HandIII、EcoR I 和标准DNA 分子( Promega 公司) 。 方法：分离、培养新西兰大白兔骨髓间充质干细胞。构建并鉴定pcDNA3.0-VEGF165 真核表达载体，利用脂质体介导其转染骨髓间充质干细胞，采用ELISA 方法检测转基因骨髓间充质干细胞的人血管内皮生长因子蛋白表达，MTT 法检测转基因骨髓间充质干细胞表达物对血管内皮细胞增殖活性的影响，设单纯培养骨髓间充质干细胞及pcDNA3.0 转染骨髓间充质干细胞组为对照组。 主要观察指标：①重组质粒双酶切和基因测序分析。②ABC-ELISA 法观察质粒转染后的骨髓间充质干细胞的人血管内皮细胞生长因子蛋白表达。③通过MTT 法检测人血管内皮生长因子165 基因转染骨髓间充质干细胞培养上清对血管内皮细胞增殖的影响。 结果：①构建的质粒用Hind III 和Xho I 双酶切、琼脂糖凝胶电泳鉴定结果与预期完全相同，PCR 和酶切鉴定正确后行测序鉴定，测序结果正确，证明所构建的质粒为pcDNA3.0-VEGF165 重组质粒。②ABC-ELISA 法检测结果表明，人血管内皮生长因子165 基因重组载体转染骨髓间充质干细胞后24 h，即有较高水平的人血管内皮生长因子165 蛋白表达，48 及72 h 呈下降趋势，但与对照组比较，差异显著( P < 0.05) 。③MTT 法检测人血管内皮生长因子165 基因转染骨髓间充质干细胞培养上清对血管内皮细胞增殖的影响，结果显示，含2%, 4%, 8%, 16%和32%转染人血管内皮生长因子165 基因的骨髓间充质干细胞培养上清促血管内皮细胞增殖率均高于对照组 ( P < 0.05) 。 结论：人血管内皮生长因子165基因可成功地转染至骨髓间充质干细胞中，并可进行有效地表达。     

构建稳定表达血管内皮细胞生长因子 的人间充质干细胞

背景：血管内皮细胞生长因子可有效治疗缺血性心脏病,但其在体内难以保持持续的有效浓度。 目的：构建稳定表达血管内皮细胞生长因子的人骨髓间充质干细胞株,检测经基因转染后外源基因的表达。 设计、时间及地点：观察实验,于2006-03/2007-04在上海胸科医院完成。 材料：人骨髓间充质干细胞株、血管内皮细胞生长因子由上海市胸科医院胸部肿瘤研究所基础实验室提供。 方法：扩增血管内皮细胞生长因子片断,构建pcPGK- hVEGF 165真核表达质粒,利用脂质体介导转染人骨髓间充质干细胞,然后进行阳性细胞克隆筛选。 主要观察指标：以RT-PCR、PCR、Western Blot、 ELISA 方法检测在稳定转染了pcPGK-VEGF165-IRES-GFP的3株细胞和稳定转染pcPGK- IRES-GFP的3株细胞中,人血管内皮细胞生长因子165在人骨髓间充质干细胞中的表达情况。 结果：扩增血管内皮细胞生长因子后,成功了构建质粒 pcPGK-hVEGF165,并利用脂质体顺利转染了人骨髓间充质干细胞,并筛选出稳定表达的细胞株。RT-PCR、PCR检测结果显示,在稳定转染血管内皮细胞生长因子骨髓间充质干细胞中表达的血管内皮细胞生长因子165信使RNA明显高于对照及未转染的人骨髓间充质干细胞,Western Blot、ELISA检测结果显示,稳定转染血管内皮细胞生长因子人骨髓间充质干细胞及培养上清中的血管内皮细胞生长因子蛋白明显高于对照及未转染的人骨髓间充质干细胞。 结论：采用脂质体介导基因转移技术可将人血管内皮细胞生长因子165顺利转染人骨髓间充质干细胞中,并获得稳定表达血管内皮细胞生长因子165的细胞株。     

脂质体介导pcDNA3/hVEGF165转染骨髓基质干细胞复合冻干骨的体内成骨和血管化☆

背景：以往的研究表明血管内皮生长因子、碱性成纤维细胞生长因子可以促进移植物的存活和体内生长。 目的：观察脂质体介导的pcDNA3/hVEGF165转染骨髓基质干细胞后复合冻干松质骨在体内的成骨和血管化效果。 方法：取同种异体新西兰大白兔的耾骨和股骨制备冻干骨,用脂质体将血管内皮生长因子转染入体外培养扩增新西兰大白兔骨髓基质干细胞中,使其附着于同种异体冻干松质骨。将新西兰大白兔分为3组,于兔竖脊肌分别植入单纯冻干骨、单纯骨髓间充质干细胞复合冻干骨组、转染有血管内皮生长因子的骨髓基质干细胞复合冻干松质骨。 结果与结论：植入后8周时可见到转染血管内皮细胞生长因子的骨髓基质干细胞复合冻干松质骨标本的表面有软骨和骨质形成,有成骨和破骨细胞出现,并形成类骨质,在移植骨周围组织中有大量血管形成,其他两组仅有大量纤维包裹。说明,脂质体介导的pcDNA3/hVEGF165转染后骨髓基质干细胞复合冻干骨具有较好成骨活性,优于单纯骨髓基质干细胞复合冻干松质骨和单纯冻干骨。     

重组合异种骨复合血管内皮生长因子基因转染骨髓间充质干细胞对非创伤性股骨头坏死的修复*

目的：将复合有血管内皮生长因子基因转染的骨髓间充质干细胞的重组合异种骨植入病灶清除区,观察其修复股骨头坏死的效果。 方法：体外分离、培养、鉴定骨髓间充质干细胞,PcDNA3/VEGF165质粒转染骨髓间充质干细胞,与重组合异种骨复合培养。应用局部液氮冷冻法造成的24只兔非创伤性股骨头坏死模型,左侧骨缺损处植入复合血管内皮生长因子基因转染骨髓间充质干细胞的重组合异种骨为治疗组,右侧仅植入单纯重组异种骨为对照组。行X射线,组织病理及股骨骨密度和矿物质含量检查。 结果：①X射线片观察第6周时治疗组坏死区出现骨小梁结构,新骨形成增加,对照组仍呈现坏死区中心低密度,边缘高密度;第12周时治疗组股骨头形态规则,密度均匀,对照组股骨头坏死区有囊状低密度区。②第6周和12周时两组骨密度和矿物质含量差异有显著性意义(P < 0.01)。③在组织学上,6周时治疗组骨细胞及微毛细血管明显多于对照组;12周治疗组新生骨组织修复达软骨下,对照组植入骨块仍有少量未吸收。 结论：重组合异种骨复合血管内皮生长因子基因转染骨髓间充质干细胞具有明显的成骨诱导作用,能有效促进非创伤性股骨头坏死的早期修复。 关键词：股骨头坏死;骨髓间充质干细胞;血管内皮生长因子;重组合异种骨     

SOX-9和胰岛素样生长因子1共同转染大鼠骨髓间充质干细胞

背景：单基因诱导虽能对骨髓间充质干细胞向软骨细胞分化产生积极影响,但作用有限,多种基因共同作用才符合机体真实内环境的需求,并有助于发挥多基因产物之间的协同作用,提高分化效果。 目的：验证应用SOX-9和胰岛素样生长因子1基因转染大鼠骨髓间充质干细胞后,目的基因分泌情况及向软骨细胞的分化效果。 设计、时间及地点：两样本观察,实验于2008-04/2009-02在中国医科大学细胞生物实验室完成。 对象：6周龄健康雄性Wistar大鼠2只用于骨髓间充质干细胞提取。 方法：扩增、提取质粒pcDNA3.1-IGF-1、pcDNA3.1-SOX-9。分离、纯化Wistar大鼠骨髓间充质干细胞。按转染情况分为5 组：①未转染组：仅加入双无(无血清无双抗)L-DMEM。②转染空载体组：双无L-DMEM +pcDNA3.1空载体和脂质体。③转染SOX-9组：双无L-DMEM+pcDNA3.1- SOX-9质粒和脂质体。④转染胰岛素样生长因子1组：双无L-DMEM分别+pcDNA3.1-IGF-1质粒和脂质体。⑤共转染组：双无L-DMEM分别+pcDNA3.1-IGF-1、pcDNA3.1-SOX-9质粒和脂质体,混合。 主要观察指标：对转染后细胞进行筛选,MTT法测定筛选后细胞的增殖活性,反转录-聚合酶链反应和Western blot法检测筛选后细胞目的基因和蛋白的表达。 结果：MTT法检测转染SOX-9组、转染胰岛素样生长因子1组和共转染组骨髓间充质干细胞增殖活性A值显著高于未转染组、转染空载体组(P < 0 01)。反转录-聚合酶链反应和Western blot检测目结果显示,SOX-9表达以转染SOX-9组最多;胰岛素样生长因子1表达以共转染组最多;软骨细胞特异性基质Ⅱ型胶原表达以共转染组最多。 结论：双基因转染在诱导骨髓间充质干细胞向软骨细胞转化的能力和细胞外基质分泌的能力上更明显高于单基质转染;另外结果间接说明胰岛素样生长因子1具有诱导骨髓间充质干细胞向软骨细胞分化的作用,但能力弱于SOX-9。     

转基因骨髓间充质干细胞移植与组织工程皮肤的血管化

背景：组织工程化皮肤移植后常因缺乏足够的血管化而导致低灌注和缺血损伤。利用转基因技术,可将血管生成调控因子基因导入目的细胞,使其表达某些调控因子,进而利于血管生成。 目的：观察转基因骨髓间充质干细胞移植促进组织工程皮肤血管化基因治疗的可行性。 设计、时间及地点：随机对照动物实验,于2008-03/11在江西省南昌大学第一附属医院烧伤研究所完成。 材料：人骨髓间充质干细胞由试验者取自临床骨髓穿刺检查骨髓正常的患者。pShuttle-CMV/VEGF165 质粒转化大肠杆菌菌种由武汉协和医院郜勇博士惠赠。16只新西兰大白兔用于皮肤缺损模型的制作,分为基因转染骨髓间充质干细胞组、骨髓间充质干细胞组、真皮支架材料组进行移植。 方法: 采用密度梯度离心结合贴壁法分离培养人骨髓间充质干细胞,以pShuttle-CMV/VEGF165质粒转染骨髓间充质干细胞。于兔背部两侧制备2 cm×2 cm 的正方形全层皮肤缺损3个,并分别以人血管内皮细胞生长因子165转染骨髓间充质干细胞、骨髓间充质干细胞、不含细胞的真皮支架材料植入全层皮肤缺损创面。 主要观察指标：观察局部组织微血管密度变化及移植物成活率。 结果：术后7 d,3组创口周围皮肤均无明显红肿及炎症反应,移植物与创面接触紧密,基因转染骨髓间充质干细胞组、骨髓间充质干细胞组可见部分真皮泛红, 真皮支架材料组不明显,术后3周创面基本愈合,基因转染骨髓间充质干细胞组创面的毛细血管密度较骨髓间充质干细胞组、真皮支架材料组明显增高(P < 0.01),14 d时3组中基因转染骨髓间充质干细胞组血管密度仍最高,但3组数据无统计学差异。术后二三周基因转染骨髓间充质干细胞组移植物成活率最高(P < 0.01)。 结论：血管内皮细胞生长因子转染骨髓间充质干细胞移植可改善和促进局部血管再生,促使新的毛细血管从创面长入移植的组织工程皮肤,从而促进组织工程皮肤的早期血管化及创面愈合。     

血管内皮细胞生长因子/骨形态发生蛋白2联合修饰骨髓间充质干细胞修复股骨头坏死

背景：将血管内皮细胞生长因子及骨形态发生蛋白二者联合修复坏死骨,有可能在保持种子细胞成骨表型的同时,又能持续高效地分泌血管内皮细胞生长因子及骨形态发生蛋白2,从而有效促进血管再生,促进组织工程化骨组织的形成和再血管化。 目的：观察血管内皮细胞生长因子165/骨形态发生蛋白2体外修饰骨髓间充质干细胞移植治疗兔股骨头缺血性坏死效果。 方法：建立兔右侧股骨头缺血性坏死模型,并以抽签法随机分为3 组：单纯髓芯减压组、髓芯减压+骨髓间充质干细胞组、髓芯减压+血管内皮细胞生长因子165/骨形态发生蛋白2转染骨髓间充质干细胞组。关节镜监视下将坏死骨清除,组织学检查成骨及血管生成情况。 结果与结论：通过关节镜观察显示各组坏死骨清除干净,有新鲜血流出。组织形态学检测发现,髓芯减压+血管内皮细胞生长因子165/骨形态发生蛋白2转染骨髓间充质干细胞组移植后不同时间点血管数量及修复区新骨面积比显著高于单纯髓芯减压组、髓芯减压+骨髓间充质干细胞组(P < 0.05)。提示血管内皮细胞生长因子165/骨形态发生蛋白2基因转染加强了骨髓间充质干细胞成骨作用,提高了新生骨的数量与质量,加快了股骨头缺血性坏死的修复。     

体外培养条件下人骨髓间充质干细胞碱性成纤维细胞生长因子基因的转染与鉴定

背景：骨髓间充质干细胞的定向分化需要合适生长因子的调控,利用细胞因子基因转染促进其生长、定向分化和加速骨缺损修复是近年来研究热点。 目的：探讨在体外培养条件下转染碱性成纤维细胞生长因子基因对人骨髓间充质干细胞生物学特性的影响。 方法：将含人pcDNA3.1-bFGF真核表达载体DH-5α菌扩增,用EndoFree质粒抽提纯化试剂盒抽提质粒,并对所提取的pcDNA3.1-bFGF重组表达质粒进行酶切鉴定和测序。利用脂质体将pcDNA3.1-bFGF质粒转染到生长良好的P3代骨髓间充质干细胞,G418筛选获得抗性克隆。采用荧光定量PCR、免疫组化、免疫荧光、Western-blot检测转染后骨髓间充质干细胞碱性成纤维细胞生长因子基因及其产物的表达,流式细胞仪检测细胞增殖周期。 结果与结论：脂质体介导pcDNA3.1-bFGF重组表达质粒转染骨髓间充质干细胞后,转染细胞确实表达碱性成纤维细胞生长因子,且表达部位主要位于胞浆。转染细胞增殖活力加强,处于增殖周期的细胞比例更高(P < 0.05)。提示采用脂质体转染法能将pcDNA3.1-bFGF成功导入体外培养的骨髓间充质干细胞,碱性成纤维细胞生长因子基因转染后可改善骨髓间充质干细胞的生存状态,促进其增殖。     

碱性成纤维细胞生长因子基因转染兔骨髓间充质干细胞*☆

背景：碱性成纤维细胞生长因子对于创伤修复具有明显的促进作用,但局部应用难以持久地发挥作用。 目的：观察碱性成纤维细胞生长因子基因转染至兔骨髓间充质干细胞后的表达情况。 设计、时间及地点：细胞基因学体外观察,于2009-03/08在云南大学生命科学院完成。 材料：日本大耳白兔1只,购于昆明医学院动物科。pCDNA3.1质粒为美国Invitrogen产品。 方法：抽取兔髂前上棘骨髓,采用贴壁法分离培养骨髓间充质干细胞,待细胞融合至培养瓶底80%时消化传代。参考GeneBank碱性成纤维细胞生长因子cDNA序列设计合成基因,电泳纯化后xholⅠ、BamHⅠ双酶切回收凝胶,以限制性核酸内切酶对PcDNA Vector质粒进行酶切、电泳和凝胶回收,连接碱性成纤维细胞生长因子DNA与PcDNA Vector质粒,构建pCDNA-碱性成纤维细胞生长因子真核表达载体,运用脂质体转染法将重组体转染至兔骨髓间充质干细胞中,并筛选稳定转染株。 主要观察指标：骨髓间充质干细胞表面抗原的表达,Western blot法检测目的蛋白的表达。 结果：流式细胞仪检测结果示所培养细胞表面抗原呈CD90,CD44阳性,CD45呈阴性;免疫组化检测结果示骨髓间充质干细胞血管源性细胞表面抗原CD34呈阴性,而相对特异性标记物CD44呈阳性。转染碱性成纤维细胞生长因子的骨髓间充质干细胞提取的细胞裂解液中,在Mr 23 000处有一条明显阳性杂交带;而转染pCDNA3.1(-)空载体的骨髓间充质干细胞提取蛋白未见阳性条带。 结论：实验成功地将碱性成纤维细胞生长因子基因转染至兔骨髓间充质干细胞,该目的基因可稳定表达。     

重组碱性成纤维细胞生长因子基因转染对于骨髓源性成骨细胞的

背景：碱性成纤维细胞生长因子对来源于中胚层和神经外胚层的细胞具有明显的促进增殖作用,对骨髓间充质干细胞具有间接的成骨作用。 目的：观察碱性成纤维细胞生长因子基因转染对骨髓源性成骨细胞生物学特性和血管生成的影响,与骨髓间充质干细胞作对比。 设计、时间及地点：对比分析基因转染效果实验,于2007-11-27/2008-12-01在南方医科大学珠江医院中心实验室和动物实验中心完成。 材料：2月龄新西兰大白兔用于骨髓间充质干细胞的分离培养及成骨细胞诱导。5周龄Wistar大鼠用于移植实验。牛pcDNA3-bFGF真核表达质粒由中山大学中山医学院免疫教研室徐霖博士惠赠。 方法：分离兔骨髓间充质干细胞和骨髓源性成骨细胞,采用脂质体介导将牛pcDNA3-bFGF真核表达质粒分别转染体外培养的兔骨髓间充质干细胞和诱导的骨髓源性成骨细胞,G418筛选。稳定转染碱性成纤维细胞生长因子的骨髓源性成骨细胞与磷酸钙胶原材料复合培养,植于Wistar大鼠后腿肌袋内,正常饲养2,4周后取出植入的复合材料,观察血管新生情况。 主要观察指标：碱性成纤维细胞生长因子转染前后骨髓间充质干细胞和骨髓源性成骨细胞生物学特性参数的比较以及骨髓源性成骨细胞转染碱性成纤维细胞生长因子前后新生血管数目的比较。 结果：利用RT-PCR、免疫细胞化学染色分析转染细胞的碱性成纤维细胞生长因子表达情况。从形态、增殖特性和细胞周期、碱性磷酸酶、Ⅰ型胶原等方面分析稳定表达碱性成纤维细胞生长因子的骨髓间充质干细胞和骨髓源性成骨细胞生物学特性。重组牛pcDNA3-bFGF真核表达质粒成功转染目的细胞,经G418筛选稳定表达。转染的细胞有大量目的基因mRNA转录和目的蛋白表达。转染碱性成纤维细胞生长因子基因的骨髓间充质干细胞和骨髓源性成骨细胞生长均加快,细胞周期中增殖期细胞比例明显增加,细胞形态无明显变化。转染与未转染的骨髓源性成骨细胞比较,碱性磷酸酶的分泌量无明显变化(P > 0.05),转染后骨髓间充质干细胞的碱性磷酸酶分泌量小于骨髓源性成骨细胞(P < 0.01)。骨髓源性成骨细胞转染与未转染碱性成纤维细胞生长因子基因组均有Ⅰ型胶原mRNA表达,但是,转染和未转染的骨髓间充质干细胞中均无Ⅰ型胶原基因mRNA转录。转染有外源性碱性成纤维细胞生长因子基因的复合材料部分被增生纤维结缔组织包绕,内有新生血管生成,随着时间的延长而增多。 结论：稳定表达碱性成纤维细胞生长因子的骨髓间充质干细胞和骨髓源性成骨细胞增殖较快,但作为组织工程骨的种子细胞,骨髓源性成骨细胞较骨髓间充质干细胞更理想。外源碱性成纤维细胞生长因子基因转染后稳定表达,对新生血管的形成具有促进作用,可以作为组织工程化人工骨种子细胞转染的目的基因。     

Epilepsy and anomalies of neuronal migration: MRI and clinical aspects

Neuronal migration disorders are the result of disturbed brain development. In such disorders, neurons are abnormally located. In diagnosing these conditions, magnetic resonance imaging is superior to any other imaging technique. This enables us to improve our knowledge of the clinical correlates of neuronal migration. With reference to migrational disorder, a retrospective study of all 303 patients with epileptic seizures referred for magnetic resonance imaging during a 3-year period was performed, 13 patients (aged 12-41, mean age 27) were identified. They represent 4.3% of the entire study group. Of the patients with known epilepsy, 6.7% and of the mentally retarded, 13.7% had migrational disorders. Four patients had schizencephaly as the dominant finding, one was classified as hemimegalencephaly, 2 had isolated heterotopias, and 6 had localized pachy- and/or poly-microgyria. The clinical pictures are complex. Ectopias of grey matter are recognised foci of epilepsy, but from an epileptological and a clinical viewpoint little attention has been given to these disorders. The present study shows that malmigration is not rare in epilepsy patients, especially not in the mentally retarded.     

Hepatic Considerations in the Use of Antiepileptic Drugs

Summary: Virtually all of the major antiepileptic drugs (AEDs) can cause hepatotoxicity, although fatal hepatic reactions are rare. The mechanisms, incidences, and risk profiles for such reactions differ from drug to drug. With carbamazepine and phenytoin, hepatotoxicity may be due to drug hypersensitivity. Although the profiles of patients at risk have not been well-defined for these two antiepileptic drugs, it would appear from reports in the literature that older adolescents and adults are at higher risk than children of developing serious or fatal hepatotoxicity. Once hepatotoxicity develops, mortality rates are 10–38% with phenytoin and 25% for carbamazepine. The risk profile for valproate fatal hepatotoxicity has been more clearly defined. Those at primary risk of fatal hepatic dysfunction are children under the age of 2 years who are receiving multiple anticonvulsants and also have significant medical problems in addition to severe epilepsy. The risk is considerably lower for patients over the age of 2 years on valproate monotherapy. In contrast to the risk profile with other AEDs, adults receiving valproate as monotherapy have the lowest risk of hepatotoxicity. Fatal hepatic dysfunction coincident with valproate may be the result of aberrant drug metabolism. Concomitant use of AEDs that induce microsomal P450 enzymes (e.g., phenytoin and phenobarbital) may enhance the production of a toxic metabolite, and hence the greater risk of hepatotoxicity with polypharmacy.     

Vascular Malformations and Epilepsy: Clinical Considerations and Basic Mechanisms

Summary: Vascular malformations (VMs) are associated with epilepsy. The natural history of the various VMs, clinical presentation, and tendency to provoke epilepsy determine treatment strategies. Investigations have probed the mechanisms of epileptogenesis associated with these lesions. Electrophysiologic changes are associated with epileptogenic cortex adjacent to VMs. Putative pathophysiologic mechanisms of epileptogenesis include neuronal cell loss, glial proliferation and abnormal glial physiology, altered neurotransmitter levels, free radical formation, and aberrant second messenger physiology.     

Classification of methods in transcranial Electrical Stimulation (tES) and evolving strategy from historical approaches to contemporary innovations

Transcranial Electrical Stimulation (tES) encompasses all methods of non-invasive current application to the brain used in research and clinical practice. We present the first comprehensive and technical review, explaining the evolution of tES in both terminology and dosage over the past 100 years of research to present day. Current transcranial Pulsed Current Stimulation (tPCS) approaches such as Cranial Electrotherapy Stimulation (CES) descended from Electrosleep (ES) through Cranial Electro-stimulation Therapy (CET), Transcerebral Electrotherapy (TCET), and NeuroElectric Therapy (NET) while others like Transcutaneous Cranial Electrical Stimulation (TCES) descended from Electroanesthesia (EA) through Limoge, and Interferential Stimulation. Prior to a contemporary resurgence in interest, variations of transcranial Direct Current Stimulation were explored intermittently, including Polarizing current, Galvanic Vestibular Stimulation (GVS), and Transcranial Micropolarization. The development of these approaches alongside Electroconvulsive Therapy (ECT) and pharmacological developments are considered. Both the roots and unique features of contemporary approaches such as transcranial Alternating Current Stimulation (tACS) and transcranial Random Noise Stimulation (tRNS) are discussed. Trends and incremental developments in electrode montage and waveform spanning decades are presented leading to the present day. Commercial devices, seminal conferences, and regulatory decisions are noted. We conclude with six rules on how increasing medical and technological sophistication may now be leveraged for broader success and adoption of tES.     

Carbamazepine Efficacy and Utilization in Children

Summary: Carbamazepine is effective for preventing partial and generalized tonic-clonic seizures in children. Although absence epilepsies are more common in children than adults, an estimated 80% of children with epilepsy have seizure types or epilepsies that are potentially responsive to carbamazepine. The differential diagnosis of ictal staring is an especially important issue in children because absence and atypical absence seizures are more prevalent in children than adults. Age-related pharmacokinetic differences and drug interactions are major considerations in children. On average, children have higher clearance rates of carbamazepine, shorter half-lives, and higher ratios of carbamazepine-10, 11-epoxide to carbamazepine than adults. In addition, children with severe epilepsy are more likely to require multiple-drug therapy, which can lead to complex drug interactions. When carbamazepine is administered along with valproate, drug protein binding interactions can cause intermittent side effects.     

Neonatal Seizures: Problems in Diagnosis and Classification

Summary: The clinical identification of neonatal seizures is critical for the recognition of brain dysfunction; however, diagnosis is often difficult because of the poorly organized and varied nature of these behaviors. Current classification systems are limited in their ability to communicate motor, autonomic, and electroencephalo-graphic features of seizures precisely and to provide a basis for uniform effective diagnosis, therapy, and determination of prognosis. Recent investigations of neonates, utilizing bedside electroencephalographic/polygraphic/ video monitoring techniques, have provided the basis for improved diagnosis and classification of seizures in the newborn. These studies have demonstrated that not all clinical phenomena currently considered to be seizures require electrocortical epileptiform activity for their initiation or elaboration. In addition, the specific clinical character of the phenomena considered to be seizures, the clinical state of the infant, and the character of the EEG indicate the probable pathophysiological mechanisms involved and suggest probable etiologies, prognosis, and therapy. Similarities between animal models that demonstrate reflex physiology and neonates with motor automatisms and tonic posturing suggest that these clinical behaviors may not be epileptic in origin but, rather, primitive movements of progression and posture mediated by brainstem mechanisms. Although not all clinical behaviors currently considered to be neonatal seizures may have similar pathophysiological mechanisms, they are clinically significant because they all indicate brain dysfunction.     

Valproate Monotherapy in the Management of Generalized and Partial Seizures

Summary: For decades, therapeutic tradition has promoted the concept of polypharmacy in the management of epilepsy. In recent years, however, studies have shown that, for most patients, monotherapy can provide comparable or better seizure control than administration of multiple anticonvulsants, while diminishing the potential for adverse reactions, drug interactions, and poor compliance. Valproate is an important monotherapeutic agent that is highly effective in the control of idiopathic primary and secondarily generalized epilepsies, and partial seizures that do not generalize. Comparative studies have found that valproate is at least as effective as phenytoin and carbamazepine in the treatment of generalized and partial seizures. Given the similar efficacy, other factors such as pharmacokinetics and side effects may therefore determine anticonvulsant selection for monotherapy.     

Un nouveau cadre conceptuel de travail pour la psychiatrie

In an attempt to place psychiatric thinking and the training of future psychiatrists more centrally into the context of modern biology, the author outlines the beginnings of a new intellectual framework for psychiatry that derives from current biological thinking about the relationship of mind to brain. The purpose of this framework is twofold. First, it is designed to emphasize that the professional requirements for future psychiatrists will demand a greater knowledge of the structure and functioning of the brain than is currently available in most training programs. Second, it is designed to illustrate that the unique domain which psychiatry occupies within academic medicine, the analysis of the interaction between social and biological determinants of behavior, can best be studied by also having a full understanding of the biological components of behavior.